RT—PCR检测MDR1基因在白血病患者中的表达

应用ＲＴ－ＰＣＲ方法建立了定量检测ＭＤＲ１表达水平的方法，并对１６５例２０５份血液及骨髓细胞标本进行了检测，结果表明了ＲＴ－ＰＣＲ方法是一项敏感，特异及适于临床的检测手术。     

结直肠癌组织缺陷基因的杂合性缺失及其意义

目的：探讨结直肠癌组织缺陷基因DCC的缺失与结直肠癌临床病理特征之间的关系。方法：采用聚合酶链反应方法(polymerase chain reaction，PCR)对23例结直肠癌组织新鲜标本DCC基因杂合性缺失(loss of heterozygosity，LOH)进行研究。结果：1)结直肠癌患者标本中18例为杂合子，其中7例存在DCC基因杂合缺失情况；2)其DCC杂合缺失与肿瘤淋巴结转移的情况关系密切，有淋巴结转移的患者DCC的杂合缺失为6例，占所有存在杂合病例数的85．7％。结论：DCC基因的杂合缺失可能对肿瘤的转移起促进作用。     

结直肠癌组织HDAC1 mRNA表达临床意义的探讨

目的:探讨HDAC1mRNA在结直肠癌组织中的表达并分析其临床意义。方法:RT-PCR检测50例结直肠癌组织、50例癌旁组织、20例正常结直肠组织和20例腺瘤组织中HDAC1mRNA表达情况,统计学分析HDAC1mRNA相对表达量与患者临床病理特征关系。结果:癌组织HDAC1mRNA相对表达量(0.608±0.032)明显高于癌旁组织(0.496±0.026),腺瘤组织HDAC1mRNA相对表达量(0.495±0.052)明显高于正常组织(0.388±0.050);癌组织中HDAC1mRNA相对表达量与患者的性别(t=0.264,P=0.793)、年龄(t=0.116,P=0.908)和肿瘤大小(t=0.621,P=0.537)无关,而与组织Duke分期(t=3.395,P=0.001)、组织学分型(t=2.070,P=0.043)和淋巴结转移(t=2.162,P=0.036)相关。Duke C、D期HDAC1mRNA相对表达量明显高于Duke A、B期,高、中分化结直肠癌组织HDAC1mRNA相对表达量明显低于低、未分化组织,淋巴结转移患者HDAC1mRNA相对表达量增高。CK20(t=2.389,P=0.021)和Ki-67(t=2.117,P=0.039)高表达患者HDAC1mRNA相对表达量明显增高。结论:HDAC1mRNA高表达与结直肠癌患者临床病理学特征相关。RT-PCR检测HDAC1mRNA在结直肠癌组织中的表达,可能成为一种新的结直肠癌特异性诊断标志。     

环氧合酶-2 mRNA在结直肠癌组织中表达的意义

流行病学调查认为非甾体类抗炎药可以降低结直肠癌死亡率，主要作用靶点是环氧合酶-2（COX-2）。COX-2作为前列腺素生物合成的限速酶，可能在结直肠肿瘤发生和发展中起到重要作用。本研究应用实时荧光定量RT-PCR方法定量分析了结直肠正常黏膜、腺瘤、癌和转移组织中COX-2 mRNA的表达水平，探讨COX-2在结直肠癌发生和发展中的意义。     

耐药基因MDR1、GST-π在乳腺癌组织中的表达及临床应用

肿瘤耐药是；临床化疗失败的主要原因，经研究发现某些肿瘤耐药是多机制的，单一检测某项指标并不全面，联合检测会提高阳性率，给临床提供更多的耐药信息。近年来，用于检测比较多的是多药耐药基因ＭＤＲ１和其编码的Ｐ－糖蛋白，ＧＳＴ－π。在多药耐药中的作用及其表达情况也成为研究热点。据报道ＭＤＲ１和ＧＳＴ－π高度相关，联合检测可明显提高耐药的检出率，本研究试图采用联合检测方法了解其表达水平，及其与临床耐药的相关性。１材料与方法１．１临床资料及试剂１．１．１ ８４例乳腺癌标本均为本院乳腺科手术提供，经乳腺病理室…     

RT-PCR方法检测大肠癌患者外周血中组织特异性mRNA的表达及其临床意义

目的：检测大肠癌患者外周血中组织特异性mRNA的表达并分析其临床意义。方法：用RT－PCR方法检测70例大肠癌患者和35例正常健康人外周血中组织特异性mRNA的表达。结果：70例大肠癌患者中有30例外周血中组织特异性mRNA表达阳性，阳性率为42．9％，与健康对照人群相比差异有显著性（P＜0．05），DukesC和Dukes D期大肠癌患者外周血中组织特异性mRNA的表达高于Dukes A期大肠癌患者（P＜0．05），在不同分化程度大肠癌患者外周血中的表达差异无显著性（P＞0．05），手术后比手术前的阳性表达率低，但差异无显著性（P＞0．05），结论：大肠癌患者外周血中组织特异性mRNA的阳性表达同肿瘤的分期有关，可作为大肠癌微转移的监测指标。     

结直肠癌组织缺陷基因的杂合性缺失及其意义

目的:探讨结直肠癌组织缺陷基因DCC的缺失与结直肠癌临床病理特征之间的关系。方法:采用聚合酶链反应方法(polymerasechainreaction,PCR)对23例结直肠癌组织新鲜标本DCC基因杂合性缺失(lossofheterozygosity,LOH)进行研究。结果:1)结直肠癌患者标本中18例为杂合子,其中7例存在DCC基因杂合缺失情况;2)其DCC杂合缺失与肿瘤淋巴结转移的情况关系密切,有淋巴结转移的患者DCC的杂合缺失为6例,占所有存在杂合病例数的85.7%。结论:DCC基因的杂合缺失可能对肿瘤的转移起促进作用。     

沉默信息调节因子1在结直肠癌组织中的表达及意义

目的 研究沉默信息调节因子1(SIRT1)在结直肠癌组织及癌旁正常组织中的表达情况及其与临床病理特征的关系.方法 分别用实时荧光定量PCR(RT-QPCR)技术及免疫组织化学(IHC)技术检测45例结直肠癌组织及30例癌旁正常组织中SIRT1在mRNA及蛋白水平上的表达情况,分析其与临床病理特征的关系.结果 SIRT1 mRNA在结直肠癌组织中的表达量较正常组织高,其中位数(四分位数间距)分别为2.488(3.447)、1.563(2.867),差异有统计学意义(Z=-2.304,P＜0.05).SIRT1蛋白水平在结直肠癌组织及正常组织中的阳性表达率分别为71.1％(32/45)和43.3％ (13/30),差异有统计学意义(x2 =5.787,P=0.029).且SIRT1的表达水平与结直肠癌的淋巴结转移(RT-QPCR:Z=-2.160,P=0.031;IHC:P =0.043)、肿瘤分期(RT-QPCR:Z=-2.411,P=0.016;IHC:P=0.008)相关,与组织分化程度可能相关(RT-QPCR:x2=10.864,P=0.004;IHC:P =0.322),而与患者的年龄、性别、肿瘤大小、大体类型、浸润深度、肿瘤部位及远处转移无关.结论 SIRT1在结直肠癌组织中呈现高表达,其可能作为一种致癌基因参与结直肠癌的发生、发展,并可能影响患者的预后.     

RASSF1A基因在乳腺癌组织中的表达及临床病理意义

目的:探讨RASSF1A基因在乳腺癌组织中的表达以及与乳腺癌临床病理特征的关系。方法:采用逆转录聚合酶链反应(RT-PCR)技术,检测49例乳腺癌组织及癌旁非肿瘤组织中RASSF1A基因的表达。应用条带密度分析软件,定量分析RT-PCR产物电泳带密度。结果:49例癌旁非肿瘤组织中RASSF1A全部表达,RASSF1A校正值(RASSF1A/β-actin)为0.6902±0.1179;49例乳腺癌组织中仅19例RASSF1A表达,校正值为0.1833±0.2441;癌组织与癌旁非肿瘤组织中RASSF1A基因表达差异有统计学意义(t=-14.199,P=0.000)。随着临床分期的增高,腋淋巴结转移率的增加,RASSF1A基因阳性表达率降低(χ2=7.201,P=0.027;χ2=3.893,P=0.048);病理组织学分级增加,RASSF1A基因呈现低表达(χ2=6.479,P=0.039);c-erbB-2蛋白表达阳性伴随RASSF1A基因的低表达(χ2=13.437,P=0.000);即RASSF1A基因高表达与临床分期、淋巴结转移、病理组织学分级、c-erbB-2蛋白表达呈负相关。而与患者年龄(χ2=0.002,P=0.962)、病理组织学类型(χ2=0.549,P=0.908)及其雌、孕激素受体(ER/PR;χ2=0.330,P=0.566/χ2=1.464,P=0.226)状况无关。结论:乳腺癌旁非肿瘤组织中RASSF1A基因表达高于癌组织,随着临床分期的增高、病理组织学分级的上升以及c-erbB-2蛋白的高表达,RASSF1A基因表达率降低。提示RASSF1A基因丢失与乳腺癌的发生、发展及乳腺肿瘤较差的生物学特性相关。     

结直肠癌及淋巴结组织CEA mRNA表达的实时荧光定量检测

目的:建立检测CEA mRNA表达水平的实时荧光定量逆转录聚合酶链反应(FQ-PCR)方法,并探讨结直肠癌及淋巴结组织中CEA mRNA表达的应用价值。方法:应用SYBR Green Ⅰ作为荧光染料,以GAPDH作为内对照,建立检测CEA mRNA的FQ-PCR方法,并对56例结直肠癌组织、淋巴结组织CEA mRNA的表达水平进行检测。结果:CEA mRNA在结直肠癌组织中均表达,但表达水平在不同病理分化程度(P=0.8661)及不同Duke分期(P=0.2986)中无明显变化;在检测的231个淋巴结中,共有137个淋巴结CEA mRNA表达为阳性,表达水平为18.31±36.85,其阳性表达率为59.3%(137/231),明显高于常规病理检测[29.4%(68/231)],χ2=41.7497,P=0.0001;CEA mRNA在Duke C D期淋巴结中表达水平(32.85±53.49)显著高于Duke A B期(6.54±14.57),P=0.0001。结论:FQ-PCR方法简便、特异和可靠;比常规病理检查更加敏感,可有效补充常规病理检查的不足,在预测肿瘤微转移方面具有重要的临床应用价值。     

Twist在涎腺腺样囊性癌中的表达及其临床意义(英文)

Objective:The purpose of this study was to investigate the expression of Twist in salivary adenoid cystic carcinoma (SACC) and the relations between Twist expression and the clinicopathological characteristics. Methods: The expression of Twist was examined immunohistochemically in 48 cases of SACC, 18 of pleomorphic adenoma and 10 of normal parotid gland. The relationship between Twist expression in SACC and the clinicopathological factors was analyzed. Results: Twist expression was significantly higher in ...     

Aberrant TIG1 methylation associated with its decreased expression and clinicopathological significance in hepatocellular carcinoma

Recently, it has been reported that tazarotene-induced gene 1 (TIG1) methylation was frequently detected in a variety of human cancers. However, the relationship between the TIG1 methylation and the characteristics of hepatocellular carcinoma (HCC) remains unknown. The aim of present study was to observe the promoter methylation of TIG1 in HCC tissues and assess its prognostic significance for HCC. Real-time quantitative polymerase chain reaction and methylation-specific polymerase chain reaction were used, respectively, to examine the mRNA expression and methylation status of TIG1 in 91 pairs of HCC and adjacent noncancerous tissues. The mRNA expression level of TIG1 was significantly lower in HCC tissues than in adjacent noncancerous tissues. The rate of TIG1 promoter methylation was significantly higher in HCC tissues than in adjacent noncancerous tissues (P?<?0.001). A strong correlation between downregulation and promoter methylation was found in these tumors (P?<?0.001). More importantly, TIG1 methylation status was related to tumor size (P?=?0.015), histological differentiation (P?=?0.004), and tumor stage (P?<?0.001). Kaplan–Meier survival analysis showed that TIG1 promoter hypermethylation was associated with a worse outcome in patients with HCC. Further, Cox multivariate analysis indicated that TIG1 methylation status was an independent prognostic factor for the overall survival rate of HCC patients. In conclusion, our data suggested that epigenetic silencing of TIG1 gene expression by promoter hypermethylation may play an important role in HCC.     

LincRNA-PVT1在甲状腺癌组织中的表达及意义

目的：研究长链非编码RNA-PVT1（large intergenic non-coding RNA-PVT1,LincRNA-PVT1）在甲状腺癌中的表达及意义。方法选取甲状腺癌组织70例,甲状腺良性结节组织50例,分别选取距离肿瘤（良性结节）边缘＞2 cm的癌旁（结节旁）组织作为对照。SYBR green实时荧光定量PCR法检测LincRNA-PVT1在甲状腺癌、癌旁、良性结节和结节旁4类组织中的相对表达量,分析甲状腺癌中LincRNA-PVT1的相对表达与临床病理特征的关联。结果甲状腺癌、癌旁、良性结节、结节旁组织中LincRNA-PVT1的相对表达量分别为3．27±0．43、2．39±0.99、2．57±0．54、2．45±0．76,甲状腺癌组织中PVT1的相对表达量高于其他三组（均P＜0．05）,其他三组间比较差异均无统计学意义（均P＞0．05）。甲状腺癌中LincRNA-PVT1的相对表达量在TNM分期方面差异具有统计学意义（P＜0．05）,而在年龄、性别、游离三碘甲原氨酸（free triiodothyronine antigen,FT3）水平方面差异均无统计学意义（均P＞0．05）。结论 LincRNA-PVT1在甲状腺癌组织中表达升高,其表达与患者TNM分期有关。     

Modulation of MDR-1 expression by a H-ras oncogene in a human colon carcinoma cell line

In an established colon differentiation model, introduction of a c-H-ras-1 oncogene into a poorly differentiated human colon carcinoma cell line (Clone A) results in changes associated with the acquisition of a more differentiated phenotype. Down-regulation of mdr-l mRNA was shown to accompany ras-related differentiation events resulting in decreased Pgp synthesis and a significant reduction in membrane Pgp as detected by immunoprecipitation, Western-blot and FACS analysis. Consistent with these observations was a reduction in Pgp-mediated drug resistance associated with Clone-A ras transfectants, with no alteration in drug sensitivity being observed with non-MDR drugs in these cells. An alternative differentiation model involves exposure of Clone-A cells to sodium butyrate. Under these conditions, differentiation-related changes resulted in up-regulation of mdr-1 mRNA and Pgp synthesis, although no alteration in drug sensitivity was recorded. In agreement with this observation, the levels of membrane-associated Pgp remained unchanged throughout the period of exposure to sodium butyrate. This study shows that modulation of Pgp expression in colon differentiation is dependent upon the differentiation induction agent used.     

CTHRC1在食管鳞癌中的表达及其临床意义

目的 检测CTHRC1在食管鳞癌组织中的表达并分析其临床意义。方法 通过组织芯片和免疫组化技术检测215例食管鳞癌组织中CTHRC1蛋白表达,分析其异常表达与临床病理特征以及预后之间的关系。结果 CTHRC1在癌旁鳞状上皮组织中基本不表达,而在食管鳞癌组织中阳性表达率为61.9％（133／215）。CTHRC1表达与食管鳞癌TNM分期（P＜0.001）、浸润深度（P=0.006）和淋巴结转移（P=0.002）有关。单因素生存分析显示食管癌CTHRC1阳性表达者预后较CTHRC1阴性表达者差（中位OS：57个月 vs. 73个月,P=0.016）;多因素回归分析显示CTHRC1表达不是食管鳞癌独立的预后因素。结论 CTHRC1在食管鳞癌的发生和发展过程中异常活化和表达,可能是食管癌治疗的潜在靶点之一。     

Overexpression of ADP-ribosylation factor 1 in human gastric carcinoma and its clinicopathological significance

Gastric cancer is the sixth leading cause of cancer-related death in Taiwan, and the identification of related factors is essential to increase patient survival. ADP-ribosylation factor 1 (ARF1) was initially identified using 2-D electrophoresis combined with MALDI-time-of-flight mass spectrometry. ADP-ribosylation factor 1 belongs to the Ras superfamily or GTP-binding protein family and has been shown to enhance cell proliferation. In the current study, we evaluated the potential of ARF1 as a biomarker for gastric cancer detection. ADP-ribosylation factor 1 mRNA was upregulated in tumor tissues (compared with adjacent non-tumor tissues, n = 55) in approximately 67.2% of gastric cancer patients. Expression of ARF1 protein was additionally observed using Western blot and immunohistochemistry (IHC) analyses. The clinicopathological correlations of ARF1 were further evaluated. Elevated ARF1 expression was strongly correlated with lymph node metastasis (P = 0.008), serosal invasion (P = 0.046), lymphatic invasion (P = 0.035), and pathological staging (P = 0.010). Moreover, the 5-year survival rate for the lower ARF1 expression group (n = 50; IHC score < 90) was higher than that of the higher expression group (n = 60; IHC score ≥ 90) (P = 0.0228, log-rank test). To establish the specific function of ARF1 in human gastric cancer, isogenic ARF1-overexpressing cell lines were prepared. Our results showed that ARF1-overexpressing clones display enhanced cell proliferation, migration, and invasion. Furthermore, ARF1-overexpression might contribute to poor prognosis of patients. These findings collectively support the utility of ARF1 as a novel prognostic marker for gastric cancer and its role in cell invasion.     

人结肠癌组织中Raf-1的表达及其临床意义

目的:研究Raf-1在人结肠癌组织中的表达与肿瘤血管生成的关系,并探讨它们的临床意义。方法:应用组织芯片技术和免疫组化法检测来自西京医院病理科2005-2006年间的87例结肠癌病例癌区、癌旁区及切缘区组织标本中Raf-1的表达,应用CD34标记的免疫组化法检测微血管密度(microvessel density,MVD);并分析Raf-1表达和MVD与结肠癌体积、转移、分化等病理特征之间的相关性。结果:Raf-1在癌组织、癌旁组织、切缘区组织的阳性率分别为86.47%、37.34%和11.03%,3者比较有显著性差异(P<0.01)。癌组织、癌旁组织、切缘区组织中MVD值分别为(61.35±11.60)、(28.34±6.70)和(8.26±3.70),3者有显著性差异(P<0.01)。Raf-1表达和MVD与肿瘤大小、转移与否密切相关。结论:Raf-1是调控人结肠癌组织血管生成的关键分子,在癌组织中Raf-1的表达与MVD值成正相关,两者与临床上肿瘤的大小、转移与否密切相关。     

促血管生成素在大肠癌组织的表达及其临床病理意义

目的　观察促血管生成素（Ａｎｇ）-１、Ａｎｇ-２及其受体Ｔｉｅ-２在大肠癌中的表达,探讨其与大肠癌血管生成的关系及临床病理意义。方法　采用免疫组化ＰＶ-９０００二步法观察５０例大肠癌和１０例正常大肠黏膜组织中Ａｎｇ-１、Ａｎｇ-２、Ｔｉｅ-２的表达,采用ＣＤ１０５标记检测微血管密度（ＭＶＤ）。结果　大肠癌组织Ａｎｇ-１的表达低于正常大肠黏膜,Ａｎｇ-２、Ｔｉｅ-２的表达高于正常大肠黏膜,差异均有统计学意义（Ｐ＜０.０５）;Ａｎｇ-１、Ａｎｇ-２的表达在不同分化程度间差异有统计学意义（Ｐ＜０.０１）;Ａｎｇ-１在淋巴结转移组的表达低于无转移组（Ｐ＜０.０５）;Ａｎｇ-２在淋巴结转移组高于无转移组（Ｐ＜０.０５）,在Ｄｕｋｅｓ'Ｃ、Ｄ期明显高于Ｄｕｋｅｓ'Ａ、Ｂ期（Ｐ＜０.０１）;Ｔｉｅ-２在淋巴结转移组的表达高于无转移组（Ｐ＜０.０５）;Ａｎｇ-２、Ｔｉｅ-２的表达与ＭＶＤ均呈正相关（ｒ＝０.３４５,Ｐ＜０.０５;ｒ＝０.２９９,Ｐ＜０.０５）。结论　促血管生成素与大肠癌的病理分化程度、Ｄｕｋｅｓ分期、淋巴结转移有关,并且在大肠癌血管生成过程中发挥重要作用。     

Vinculin在大肠癌中的表达及其临床病理意义

目的检测纽蛋白（vinculin）在大肠癌组织中的表达,并探讨其在大肠癌发生、发展中的意义。方法采用免疫荧光结合激光共聚焦显微镜对大肠正常黏膜、原发大肠癌及淋巴结转移癌组织的vinculin表达水平进行检测。结果大肠黏膜由良性向恶性转变及转移过程中,vinculin表达水平逐渐降低;有淋巴结转移的大肠癌组织vinculin表达水平明显低于无淋巴结转移者;vinculin表达水平与癌组织浸润肠壁深度呈负相关,而与癌细胞分化程度无明显相关性。结论vinculin与大肠癌的浸润转移有关,可作为大肠癌发生、发展的预测指标之一。